Northeast Asia Institute of traditional Chinese Medicine, Changchun University of Chinese Medicine, Boshuo Road, Nanguan District, Changchun, Jilin, China.
Division of Cardiovascular Medicine, Department of Medicine, Solna, Karolinska Institutet, Stockholm 171 76, Sweden.
Phytomedicine. 2024 May;127:155428. doi: 10.1016/j.phymed.2024.155428. Epub 2024 Feb 25.
Previous studies have confirmed the antioxidant and anti-inflammatory effects of active ginseng components that protect against liver injury. However, ginseng-derived nanoparticles (GDNPs), low-immunogenicity nanovesicles derived from ginseng, have not been reported to be hepatoprotective.
In this study, we investigated whether GDNPs could attenuate alcohol-induced liver injury in LO cells and mice by modulating oxidative stress and inflammatory pathways, thereby advancing the theoretical basis for the development of novel pharmacological treatments.
Alcohol was used to construct in vitro and in vivo models of alcoholic liver injury. To explore the mechanisms by which GDNPs exert their protective effects against alcoholic liver injury, we examined the expression of oxidative stress-related genes and analysed inflammatory responses in vitro and in vivo. The experimental findings were verified using network pharmacology.
The composition of the GDNPs was analysed using liquid chromatography-mass spectrometry. GDNPs were extracted and purified using differential ultracentrifugation and sucrose density gradient centrifugation. In vitro models of alcoholic liver injury were established using LO cells, whereas C57BL/6 J mice were used as in vivo models. Oxidative stress, inflammation, and liver injury indicators were measured using appropriate kits. Levels of proteins associated with oxidative stress and inflammation were measured via western blot, while nuclear factor erythroid2-related factor 2 (Nrf2) and NF-κB protein expression was tested using immunofluorescence, immunohistochemistry, and flow cytometry. The levels of relevant transcription factors were determined using qPCR. Experimental haematoxylin and eosin staining was used to characterise the liver histological appearance and damage in mice. Network pharmacological analysis of GDNP mRNA sequencing of GDNPs was used to predict drug targets and disease associations using TCMSP.
GDNPs primarily included 77 compounds, including organic acids and their derivatives, amino acids and their derivatives, sugars, terpenoids, and flavonoids. GDNPs have features that allow them to be taken up by LO cells and promote their proliferation. In vitro data indicated that GDNPs reduced the levels of alcohol-induced reactive oxygen species by activating the Nrf2/HO-1 signalling pathway, whilst inhibiting the NF-κB pathway and thereby reducing NO, tumour necrosis factor-α, and interleukin-1β levels to alleviate inflammation. An in vivo model showed that GDNPs improved the liver parameters and pathology in mice with alcoholic liver injury. GDNPs activate the Nrf2/HO-1/Keap1 signalling pathway in a p62-dependent manner to exert antioxidant effects. Furthermore, the TLR4/NF-κB signalling pathway was involved in the in vivo anti-inflammatory effect. Network pharmacology also confirmed that the effects of GDNPs on liver disease were associated with oxidative stress and inflammation-related targets and pathways.
This study showed for the first time that GDNPs can alleviate alcohol-induced liver damage by activating the Nrf2/HO1 signalling pathway and blocking the NF-κB signalling pathway, thus lowering oxidative stress and inflammatory responses. Hereby, we present the Nrf2/HO1 and NF-κB signalling pathways as potential targets and GDNPs as a novel therapeutic approach for the management of alcohol-induced liver damage.
先前的研究已经证实，人参中的活性成分具有抗氧化和抗炎作用，可预防肝损伤。然而，人参衍生的纳米颗粒（GDNP）是源自人参的低免疫原性纳米囊泡，尚未有研究报道其具有肝保护作用。
本研究旨在探讨 GDNP 是否可以通过调节氧化应激和炎症途径减轻 LO 细胞和小鼠的酒精性肝损伤，从而为新型药理治疗方法的开发提供理论依据。
使用酒精构建体外和体内酒精性肝损伤模型。为了探讨 GDNP 发挥其对酒精性肝损伤保护作用的机制，我们在体外和体内研究了氧化应激相关基因的表达和炎症反应。网络药理学验证了实验结果。
使用液相色谱-质谱联用仪分析 GDNP 的成分。使用差速超速离心和蔗糖密度梯度离心法提取和纯化 GDNP。使用 LO 细胞建立体外酒精性肝损伤模型，使用 C57BL/6 J 小鼠建立体内模型。使用适当的试剂盒测量氧化应激、炎症和肝损伤指标。通过 Western blot 测定与氧化应激和炎症相关的蛋白质水平，通过免疫荧光、免疫组织化学和流式细胞术检测核因子红细胞 2 相关因子 2（Nrf2）和 NF-κB 蛋白表达。使用 qPCR 测定相关转录因子的水平。使用实验性苏木精和伊红染色观察小鼠肝组织的形态和损伤。使用 TCMSP 对 GDNP mRNA 测序进行网络药理学分析，预测 GDNP 的药物靶点和疾病相关性。
GDNP 主要包括 77 种化合物，包括有机酸及其衍生物、氨基酸及其衍生物、糖、萜类化合物和类黄酮。GDNP 具有被 LO 细胞摄取并促进其增殖的特征。体外数据表明，GDNP 通过激活 Nrf2/HO-1 信号通路降低酒精诱导的活性氧水平，同时抑制 NF-κB 通路，从而降低 NO、肿瘤坏死因子-α和白细胞介素-1β水平，减轻炎症。体内模型表明，GDNP 改善了酒精性肝损伤小鼠的肝参数和病理。GDNP 以 p62 依赖的方式激活 Nrf2/HO-1/Keap1 信号通路发挥抗氧化作用。此外，TLR4/NF-κB 信号通路参与了体内抗炎作用。网络药理学也证实，GDNP 对肝病的作用与氧化应激和炎症相关靶点和途径有关。
本研究首次表明，GDNP 通过激活 Nrf2/HO1 信号通路和阻断 NF-κB 信号通路减轻酒精诱导的肝损伤，从而降低氧化应激和炎症反应。本研究提出 Nrf2/HO1 和 NF-κB 信号通路可能作为潜在靶点，GDNP 可能作为治疗酒精性肝损伤的新型治疗方法。
